Reboxetine Research Articles

Development of Two Green and High-Throughput Microwell Spectrometric Platforms for Determination of Reboxetine, the First FDA-Approved Selective Noradrenaline Reuptake Inhibitor Antidepressant Drug

Abstract Background Reboxetine (RBX) is the first FDA-approved antidepressant drug of the selective noradrenaline reuptake inhibitors class. There is a serious need for a convenient analytical tool for the quantitation of RBX in its dosage form. Objective This study aims toward the development and validation of two green and high-throughput microwell spectrometric platforms for the pharmaceutical analysis of RBX. Methods The two platforms, abbreviated as MW-AB and MW-FL, involved microwell-based analysis assisted with a multifunction microplate plate reader for measuring absorbance and fluorescence signals, respectively. The MW-AB and MW-FL platforms involved the formation of colored and fluorescent derivatives upon the reaction of RBX with oxidized pyrocatechol reagent (OPC) and tetracyanoquinodimethane (TCNQ), respectively. The absorbance of colored RBX-OPC derivative at 520 nm, and the fluorescence of RBX-TCNQ charge transfer complex at 283 and 484 nm for excitation and emission, respectively. The optimum conditions of both reactions were established, their molar ratios were determined, and reaction mechanisms were postulated. Results Both platforms were optimized and validated according to the guidelines of the International Council on Harmonization. The limits of quantitation were 19.6 µg/mL and 27 ng/mL for MW-AB and MW-FL, respectively. Both platforms were applied with excellent reliability to the quantitation of RBX content in Edranox® tablets and their drug uniformity. The greenness levels of both platforms were assessed by two comprehensive tools, and the results confirmed the high level of greenness for both platforms. Conclusions Both platforms involved one-step reactions, adapted microwell analysis, and simultaneous handling of large number of samples. Therefore, they have the advantages of greenness and high-throughput analysis. Highlights The proposed two platforms are valuable tools for the rapid quantitation of RBX.

New Green and High-throughput Microwell Spectrophotometric Method for Quality Control of Pharmaceutical Formulations of Selective Serotonin Reuptake Inhibitors via Microscale Formation of their N-Vinylamino-naphthoquinone Derivatives

Background: Selective serotonin reuptake inhibitors (SSRIs) are mostly prescribed for the treatment of depression. This study describes the microscale in-microwell formation of blue-colored N-vinylamino-naphthoquinone derivatives of SSRIs upon their reaction with 2,3-dichloro-1,4- naphthoquinone (DCNQ) and acetaldehyde. The reaction was subsequently utilized as a basis for the development of a new simple and sensitive microwell spectrophotometric method (MW-SPM) for the quality control of pharmaceutical formulations of four SSRIs. These SSRIs are fluoxetine (FLU), sertraline (SER), paroxetine (PAR), and reboxetine (REB). Methods: The MW-SPM procedure was performed in 96-microwell transparent plates, and the microplate reader was employed to measure the absorbances of the reaction products at their peak absorbance wavelength of 580 nm. The best conditions for the method were determined. Results: The relations showed good linearity (correlation coefficients were ≥0.9992) in the concentration range of 5 – 600 μg/mL. The limits of detection ranged from 5.20 to 15.58 μg/mL. The precision was deemed acceptable since all cases' relative standard deviation (RSD) values remained below 2.21%. Recovery experiments were conducted to confirm the accuracy of the method, yielding recovery values of at least 97.8%. The MW-SPM method was effectively utilized to analyze SSRIs in both their bulk and pharmaceutical dosage forms, exhibiting acceptable accuracy and precision. The recovery values ranged from 99.4% to 101.0% (with a margin of error of ± 0.5% to 1.6%). The results were comparable with those of the pre-validated reported methods. Four different metric tools evaluated the greenness of the proposed method, and the results proved that the method fulfills the requirements of green analytical approaches. Furthermore, the ability to handle numerous microvolume samples simultaneously in the described method provides it with a high-throughput characteristic. Conclusion: The proposed MW-SPM represents a valuable tool for an efficient analysis of SSRIs in pharmaceutical quality control units.

Anti-Candidal Activity of Reboxetine and Sertraline Antidepressants: Effects on Pre-Formed Biofilms.

Reboxetine (REB) and sertraline (SER) are antidepressants. The antifungal potential of these drugs against planktonic Candida has been recently reported with limited data about their effects on Candidal biofilms. Biofilms are self-derived extracellular matrixes produced by the microbial population that is attached to biotic surfaces, such as vaginal and oral mucosa, or abiotic surfaces, such as biomedical devices, resulting in persistent fungal infections. The commonly prescribed antifungals, azoles, are usually less effective when biofilms are formed, and most of the prescribed antifungals are only fungistatic. Therefore, the current study investigates the antifungal potentials of REB and SER, alone and in combination with fluconazole (FLC) and itraconazole (ITR) against Candidal biofilms. Using proper controls, Candida species (Candida albicans, C. albicans; Candida krusei, C. krusei; and Candida glabrata, C. glabrata) were used to form biofilms in 96-well microplates. Serial dilutions corresponding to concentrations ranging from 2 to 4096 µg/mL of the target drugs (REB, SER, FLC, ITR) were prepared and added to the plates. Impairment of the biofilm biomass and biofilm metabolic viability was detected using the crystal violet (CV) assay and 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, respectively. In the checkerboard assay, the sessile fractional inhibitory concentration index (SFICI) was calculated to evaluate the effects of drug combinations. SER was more effective in reducing the biomass than REB for C. albicans and C. glabrata, but both were equal for C. krusei. For the reduction in metabolic activity in C. albicans and C. glabrata, SER had a slight advantage over REB. In C. krusei, REB was slightly more potent. Overall, FLC and ITR were almost equal and produced more significant reductions in metabolic activity when compared to SER and REB, except for C. glabrata, where SER was almost equal to FLC. Synergism was detected between REB + FLC and REB + ITR against biofilm cells of C. albicans. Synergism was detected between REB + ITR against biofilm cells of C. krusei. Synergism was detected between REB + FLC and REB + ITR against biofilm cells of C. albicans, C. krusei, and C. glabrata. The results of the present study support the potential of SER and REB as anti-Candidal biofilm agents that are beneficial as a new antifungal to combat Candidal resistance.

Gut Microbiome Alteration after Reboxetine Administration in Type-1 Diabetic Rats.

Antidepressants are drugs commonly used in clinical settings. However, there are very limited studies on the effects of these drugs on the gut microbiota. Herein, we evaluated the effect of reboxetine (RBX), a selective norepinephrine (noradrenaline) reuptake inhibitor (NRI), on gut microbiota in both diabetic and non-diabetic rats. This is the first report of relation between reboxetine use and the gut microbiota to our knowledge. In this study, type-1 diabetes induced by using streptozotocin (STZ) and RBX was administered to diabetic rats and healthy controls for 14 days. At the end of the treatment, stool samples were collected. Following DNA extraction, amplicon libraries for the V3-V4 region were prepared and sequenced with the Illumina Miseq platform. QIIME was used for preprocessing and analysis of the data. As a result, RBX had a significant effect on gut microbiota structure and composition in diabetic and healthy rats. For example, RBX exposure had a pronounced microbial signature in both groups, with a low Firmicutes/Bacteroidetes ratio and low Lactobacillus levels. While another abundance phylum after exposure to RBX was Proteabacteria, other notable taxa in the diabetic group included Flavobacterium, Desulfovibrionaceae, Helicobacteriaceae, Campylobacterales, and Pasteurellacae when compared to the untreated group.

Antidepressants Differentially Regulate Intracellular Signaling from α1-Adrenergic Receptor Subtypes In Vitro.

Currently utilized antidepressants have limited effectiveness and frequently incur undesired effects. Most antidepressants are thought to act via the inhibition of monoamine reuptake; however, direct binding to monoaminergic receptors has been proposed to contribute to both their clinical effectiveness and their side effects, or lack thereof. Among the target receptors of antidepressants, α1‑adrenergic receptors (ARs) have been implicated in depression etiology, antidepressant action, and side effects. However, differences in the direct effects of antidepressants on signaling from the three subtypes of α1-ARs, namely, α1A-, α1B- and α1D‑ARs, have been little explored. We utilized cell lines overexpressing α1A-, α1B- or α1D-ARs to investigate the effects of the antidepressants imipramine (IMI), desipramine (DMI), mianserin (MIA), reboxetine (REB), citalopram (CIT) and fluoxetine (FLU) on noradrenaline-induced second messenger generation by those receptors. We found similar orders of inhibition at α1A-AR (IMI < DMI < CIT < MIA < REB) and α1D‑AR (IMI = DMI < CIT < MIA), while the α1B-AR subtype was the least engaged subtype and was inhibited with low potency by three drugs (MIA < IMI = DMI). In contrast to their direct antagonistic effects, prolonged incubation with IMI and DMI increased the maximal response of the α1B-AR subtype, and the CIT of both the α1A- and the α1B-ARs. Our data demonstrate a complex, subtype-specific modulation of α1-ARs by antidepressants of different groups.

Neuro-pharmacological reinstatement of ovulation and associated neurobiology in a macaque model of functional hypothalamic amenorrhoea.

What is the underlying neuropathology in a cynomolgus macaque model of functional hypothalamic amenorrhoea (FHA) and can it be normalized to restore ovulation? Anovulatory monkeys exhibited increased hypothalamic norepinephrine (NE), kisspeptin and gonadotropin-releasing hormone (GnRH) in the early follicular phase, but administration of the NE reuptake inhibitor (NRI), reboxetine (REB), restored ovulation during stress and normalized NE, kisspeptin and GnRH. Female cynomolgus macaques, like women, show individual reproductive sensitivity to modest psychosocial and metabolic stress. During stress, resilient females ovulate through two menstrual cycles whereas stress-sensitive (SS) macaques immediately cease ovulation. On Day 5 of a non-stressed menstrual cycle, resilient macaques have less NE synthesizing enzyme [dopamine β-hydroxylase (DBH)], kisspeptin and GnRH innervation of the medial basal hypothalamus but more endogenous serotonin than SS macaques. Stress increased DBH/NE, kisspeptin and GnRH but did not alter serotonin. In a longitudinal design, 27 adult (7-13 years) female cynomolgus macaques (Macaca fascicularis) with three different levels of sensitivity to stress were monitored with daily vaginal swabs and frequent serum progesterone (P) measurements. Three 90-day experimental periods called 'Cycle Sets' were monitored. A Cycle Set consisted of one ovulatory menstrual cycle without stress, and two cycles, or 60 days, with modest stress. Each Cycle Set was followed by a rest period. During a Cycle Set, individuals were either untreated (placebo) or administered escitalopram (CIT) or REB. Ultimately, half of each sensitivity group was euthanized during stress with CIT or REB treatment and the hypothalamus was obtained. Neurobiological endpoints were compared between CIT and REB treatment groups in stress resilient and SS monkeys. The monkeys were housed at the University of Pittsburgh primate facility for the duration of the experiments. Upon euthanasia, their brains and serum samples were shipped to the Oregon National Primate Research Center. The hypothalamus was examined with immunohistochemistry for the expression of DBH (a marker for NE axons), kisspeptin and GnRH. P was measured in the serum samples by radioimmunoassay. Daily administration of REB restored ovulation in 9 of 10 SS animals during stress. Of note, REB significantly increased P secretion during stress in the most sensitive group (P = 0.032), which indicates ovulation. CIT lacked efficacy. REB significantly reduced DBH/NE, kisspeptin and GnRH axon density in the hypothalamus relative to CIT treatment (P = 0.003. 0.018 and 0.0001, respectively) on Day 5 of the menstrual cycle in resilient and sensitive groups. N/A. The US FDA has not approved REB for human use, although it is used in Europe for the treatment of depression/anxiety as EdronaxTR. Whether REB could be useful for the treatment of FHA in women has not been determined. The use of an NRI to treat FHA is a novel approach and the potential reinstatement of ovulation could be straightforward compared to current treatment protocols. The underlying neurobiology provides a compelling case for treating the origin of the pathology, i.e. elevated NE, rather than circumventing the hypothalamus altogether with gonadotropins, which have associated risks such as hyperstimulation syndrome or multiple births. Portions of this study were supported by NIH grant HD062864 to C.L.B., NIH grant HD62618 to J.L.C. and C.L.B. and 1P51 OD011092 for the operation of the Oregon National Primate Research Center. There were no competing interests.

Reboxetine and hyoscine butylbromide improve upper airway function during nonrapid eye movement and suppress rapid eye movement sleep in healthy individuals.

Recent findings indicate that noradrenergic and antimuscarinic processes are crucial for sleep-related reductions in pharyngeal muscle activity. However, there are few human studies. Accordingly, this study aimed to determine if a combined noradrenergic and antimuscarinic intervention increases pharyngeal dilator muscle activity and improves airway function in sleeping humans. Genioglossus (GG) and tensor palatini electromyography (EMG), pharyngeal pressure, upper airway resistance, and breathing parameters were acquired in 10 healthy adults (5 female) during two overnight sleep studies after 4 mg of reboxetine (REB) plus 20 mg of hyoscine butylbromide (HBB) or placebo using a double-blind, placebo-controlled, randomized, cross-over design. Compared with placebo, peak and tonic GG EMG were lower (Mean ± SD: 83 ± 73 vs. 130 ± 75, p = 0.021 and 102 ± 102 vs. 147 ± 123 % wakefulness, p = 0.021, respectively) but the sleep-related reduction in tensor palatini was less (Median [25th, 75th centiles]: 53[45, 62] vs. 34[28, 38] % wakefulness, p = 0.008) with the drug combination during nonrapid eye movement (non-REM) sleep. These changes were accompanied by improved upper airway function including reduced pharyngeal pressure swings, airway resistance, respiratory load compensation, and increased breathing frequency during N2. REB and HBB significantly reduced rapid eye movement sleep compared with placebo (0.6 ± 1.1 vs. 14.5 ± 6.8 % total sleep time, p < 0.001). Contrary to our hypothesis, GG muscle activity (% wakefulness) during non-REM sleep was lower with REB and HBB. However, sleep-related reductions in tensor palatini activity were less and upper airway function improved. These findings provide mechanistic insight into the role of noradrenergic and antimuscarinic processes on upper airway function in humans and have therapeutic potential for obstructive sleep apnea. Australian New Zealand Clinical Trials Registry, https://www.anzctr.org.au, trial ID: ACTRN12616000469415.

Clinical observation of Prolift pelvic floor reconstruction in the treatment of pelvic organ prolapse

Objective To analyze the effect of Prolift pelvic floor reconstruction on pelvic organ prolapse and its influence on quality of life (P-QOL) score and pelvic floor muscle strength level. Methods From April 2014 to February 2017, 63 patients with pelvic organ prolapse were selected and randomly divided into two groups. The patients in the control group were treated with traditional vaginal hysterectomy and anterior and posterior vaginal wall repair, while the patients in the observation group were treated with Prolift pelvic floor reconstruction. The clinical effect of two methods after treatment were compared. Results The operative time of the observation group was shorter than that in the control group. The amount of intraoperative bleeding and residual urine after operation in the observation group were lower than those in the control group. The pelvic floor muscle strength level in the observation group was superior to that in the control group. Besides, the quality of life in the observation group was higher than that in the control group one year after operation. The rate of postoperative dysuria and recurrence were lower than that of the control group (P 0.05). Conclusions Prolift pelvic floor reconstruction is a simple, less invasive and safe procedure for the correction of pelvic organ prolapse. It can promote the recovery of pelvic floor muscle function, and the curative effect is better than that of traditional surgery. Key words: Pelvic organ prolapse/SU; Surgical mesh; Postoperative complications; Treatment outcome

Acute Effect of Noradrenergic Modulation on Motor Output Adjustment in Men.

To determine the role of noradrenergic modulation in the control of motor output, we compared the acute effect of reboxetine (REB), a noradrenaline reuptake inhibitor, to a placebo (PLA) on knee extensors motor performance and cortical and spinal excitability. Eleven young men took part in two randomized experiments during which they received either 8 mg of REB or a PLA. The torque produced during a maximal voluntary contraction (MVC) and its variability (i.e., coefficient of variation) during submaximal contractions ranging from 5% to 50% MVC were measured. Paired electrical (PES) and transcranial magnetic stimulation (TMS) were used to assess changes in voluntary activation during MVC, and corticospinal (motor-evoked potential (MEP)) and spinal excitability (Hoffmann (H) reflex) during contraction at 20% MVC. MVC torque and torque steadiness increased respectively by 9.5% and 24% on average in REB compared with PLA condition (P < 0.001). Voluntary activation tested by TMS and PES was greater (~3%; P < 0.05) in REB than PLA condition. The increase in voluntary activation in REB condition was significantly correlated with subjects' initial voluntary activation level when tested by TMS (r = -0.62; P = 0.048) and PES (r = -0.86; P < 0.01). The maximal amplitudes of H reflex and MEP and the slope of their recruitment curves were enhanced by REB (P < 0.05). The ratio between the TMS-induced EMG silent period and the corresponding MEP (silent period/MEP) was reduced in REB condition (P < 0.01). The present findings indicate that voluntary activation and accuracy in force control can be increased by an enhanced level of noradrenaline concentration. This improvement in motor performance is accompanied by changes located at both cortical and spinal levels.

The effects of antidepressants on inflammatory factors of frontal lobe in depression model rats

Objective To investigate the effect of antidepressants on frontal lobe inflammatory factors in depressed model rats. Methods Thirty SD rats were randomly divided into 5 groups (n=6): contorl group(Cn), depression group(Dn), citalopram group(Dx), venlafaxine group(Dw) and reboxetine group(Dr) . The open field test and sugar consumption test were performed to observe the depression behavior and the changes of FGF-1, TNF-α, IL-1 and CRP were measured in frontal lobe of rats by ELISA. Pearson linear correlation analysis was used to evaluate the correlation between behavioral score and inflammatory factors level in rats. Results (1)Compared with the Cn group, the scores of Dn, Dr, Dw, Dx group were decreased on Open field test and sugar consumption test (P<0.05). Compared with the Dn group, the scores of the Dx, Dw, Dr group were increased in Open field test and sugar consumption test (P<0.05). (2)Compared with Cn group, the levels of TNF-α, IL-1 and CRP increased in Dn group and FGF-1 level decreased (P<0.01). Compared with Dn group, the levels of FGF-1 increased in Dx ((86.54±2.56)ng/L), Dw((79.82±4.89)ng/L) and Dr ((68.50±3.61)ng/L) group, however, the levels of TNF-α decreased in Dx ((150.21±5.65)ng/L), Dw ((161.28±8.80)ng/L), Dr ((175.78±9.67)ng/L) group, and the level of IL-1 also decreased in Dx ((30.87±4.48)ng/L), Dw ((36.65±3.33)ng/L), Dr ((40.14±2.81)ng/L) group, and the levels of CRP also decreased in Dx ((374.88±14.15)ng/L), Dw ((394.21±17.03)ng/L), Dr ((414.34±10.97)ng/L) (P<0.01). (3)The behavioral score of each group was positively correlated with of FGF-1 and negatively correlated with TNF-a, IL-1 and CRP (P<0.05). Conclusion Antidepressants can reduce the level of proinflammatory factors and increase the level of anti-inflammatory factors of frontal lobe in depression model rats, suggesting that antidepressants may play an antidepressant effect by regulating the concentration of inflammatory factors. Key words: Antidepressants; Depression model rats; FGF-1; TNF-a; IL-1; C-reaction protein

Efficacy and safety of drugs for attention deficit hyperactivity disorder in children and adolescents: a network meta-analysis.

The aim of this study is to gather evidence of head-to-head double-blind randomized-controlled trials on the efficacy and safety of available treatments for attention deficit hyperactivity disorder (ADHD) in children and adolescents. A systematic review was conducted by two independent reviewers in ten electronic databases (PROSPERO register CRD42016043239). Methodological quality of included studies was evaluated according to the Jadad scale. Network meta-analyses were performed including double-blinded head-to-head trials comparing active allopathic drugs in patients (0-18 years old) diagnosed with ADHD. The results of efficacy and safety of atomoxetine (ATX), bupropion, buspirone (BSP), dexamphetamine, edivoxetine (EDX), guanfacine (GXR), lisdexamfetamine (LDX), methylphenidate (MPH), mixed amphetamine salts, modafinil, pindolol (PDL), reboxetine (RBX), selegiline, and venlafaxine were analyzed using ADDIS software v.1.16.5. Forty-eight trials were identified (n=4169 participants), of which 12 were used for efficacy analysis and 33 for safety analysis. On the CGI-I scale, the analysis revealed that MPH was more effective than ATX and GXR. For the safety outcomes, according to drug ranks, LDX was more likely to cause sleep disorders (39%) as well as loss of appetite (65%) and behavior problems such as irritability (60%). BSP (71%) and EDX (44%) caused less appetite decrease. For behavioral effects, PDL was considered safest (50%). For any adverse events, RBX (89%) was the safest alternative. The lack of head-to-head trials properly reporting outcomes of interest limited some comparisons. Network meta-analysis offered a broader overview on the available treatments for ADHD, especially for safety issues, and contributes towards evidence gathering and clinical practice decisions. A core outcome set for ADHD should be designed to guide the conduction and report of clinical trials.

Acute and chronic effects of noradrenergic enhancement on transcranial direct current stimulation-induced neuroplasticity in humans.

Chronic administration of the selective noradrenaline reuptake inhibitor (NRI) reboxetine (RBX) increased and prolonged the long-term potentiation-like plasticity induced by anodal transcranial direct current stimulation (tDCS) for over 24h. Chronic administration of RBX converted cathodal tDCS-induced long-term depression-like plasticity into facilitation for 120min. Chronic noradrenergic activity enhancement on plasticity of the human brain might partially explain the delayed therapeutic impact of selective NRIs in depression and other neuropsychiatric diseases. Noradrenaline affects cognition and motor learning processes via its impact on long-term potentiation (LTP) and depression (LTD). We aimed to explore the impact of single dose and chronic administration of the selective noradrenaline reuptake inhibitor (NRI) reboxetine (RBX) on plasticity induced by transcranial direct current stimulation (tDCS) in healthy humans via a double-blinded, placebo-controlled, randomized crossover study. Sixteen healthy volunteers received placebo or single dose RBX (8mg) before anodal or cathodal tDCS of the primary motor cortex. Afterwards, the same subjects took RBX (8mgday-1 ) consecutively for 21days. During this period, two additional interventions were performed (RBX with anodal or cathodal tDCS), to explore the impact of chronic RBX treatment on plasticity. Plasticity was monitored by motor-evoked potential amplitudes elicited by transcranial magnetic stimulation. Chronic administration of RBX increased and prolonged the LTP-like plasticity induced by anodal tDCS for over 24h. Chronic RBX significantly converted cathodal tDCS-induced LTD-like plasticity into facilitation, as compared to the single dose condition, for 120min after stimulation. The results show a prominent impact of chronic noradrenergic enhancement on plasticity of the human brain that might partially explain the delayed therapeutic impact of selective NRIs in depression and other neuropsychiatric diseases.

Noradrenaline Reuptake Inhibition Impairs Cortical Output and Limits Endurance Time.

To assess the neural mechanisms that limit endurance time, we compared a fatiguing task performed under the influence of reboxetine (REB), a noradrenaline reuptake inhibitor, and placebo (PLA). Nine male subjects (age = 24 ± 2 yr) participated in this study. The fatiguing task involved repeated 3-s submaximal isometric contractions of the knee extensors (∼33% maximal voluntary contraction) with a 2-s rest between each contraction and performed until task failure. Before, during, and after exercise, changes in voluntary activation, corticospinal (motor-evoked potential) and spinal excitability (Hoffman reflex), and muscle contractile properties were tested using electrical and transcranial magnetic stimulations. A psychomotor vigilance task assessed reaction time before and after exercise. Compared with PLA, REB reduced the endurance time by 15.6% (P = 0.04). The maximal voluntary contraction torque decreased to a similar extent at task failure in both conditions (P < 0.01), whereas the rate of decline was greater in REB than that in PLA (P = 0.02). The level of voluntary activation tested by transcranial magnetic stimulation and electrical stimulation decreased (P < 0.01) by 10%-15% at the end of the task, but the mean rate of decline was greater in REB (P ≤ 0.03). Although motor-evoked potential did not change during fatigue, Hoffman reflex, and electrically evoked torque decreased similarly in the PLA and REB conditions (P ≤ 0.02). After exercise, reaction time increased by 3.5% (P = 0.02) in REB but did not change in the PLA condition. These findings suggest that because of the noradrenaline reuptake inhibition, the output from the motor cortex is decreased at a greater rate than that in the PLA condition, contributing thereby to shorten endurance time.

Effects of antidepressant therapy on serotonin and tryptophan hydroxylase 2 in ovariectomized rats with depressive disorder

Obiective To investigate the effects of antidepressant therapy on whole regulating function of serotonin and tryptophan hydroxylase 2 system in ovariectomized(OVX) rats with depressive disorder. Methods seventy-two female Sprague-Dawley rats were used as subjects.The rats were randomly divided into 8 groups as following: Sham operation group (Sn), depression group(Dn), the OVX group(On), OVX with depression group(ODn), OVX with depression+ sertraline group(ODs), OVX with depression+ citalopram group(ODx), OVX with depression+ venlafaxine group(ODw) , OVX with depression+ reboxetine group(ODr). Rats in group OVX and group OVX with depression were ovariectomized.Rats in group with depression were exposed to the chronic unpredictable mild stress (CUMS). Open-field test and sugar consumption experiment were tested on all rats before the experiment and the 28th day of experiment.The expression of TPH2 mRNA was measured with RT-PCR, and the expression of TPH2 protein was detected by Western blot.The concentration of 5-HT was detected by ELISA. Results (1) After exposure of CUMS, the scores of motion and percentage of sugar consumption experiment in Dn group were decreased contrast to the Sn group(P<0.05) , and the scores in ODn group were decreased compared with the on group(P<0.05). (2)The protein and gene expression of THP2 in On and Dn group were decreased contrast to the Sn group (P<0.05). The protein and gene expression of THP2 in ODn were decreased compared with the on group (P<0.05). The protein and gene expression of THP2 in ODs(protein (0.68±0.07); gene (0.87±0.09)), ODx(protein (0.71±0.05); gene (0.84±0.17)), ODw(protein (0.69±0.04); gene (0.88±0.16)) and ODr(protein (0.53±0.07); gene (0.70±0.15)) were increased compared with the ODn(protein (0.31±0.09); gene (0.56±0.16)) group(P<0.05). (3)The concentration of 5-HT of hippocampus and serum in Dn and On group were decreased contrast to the Sn group (P<0.05). The concentration of 5-HT of hippocampus and serum in ODs, ODx, ODw and ODr group was increased compared with the ODn group (P<0.05). Conclusion The antidepressants can up-regulate the expression of THP2 and the concentration of 5-HT in the OVX rats with depressive disorder.Sertralin is stronger however Reboxetine Mesylate is the poorest. Key words: Antidepressant; Depressive disorder rats model; Ovariectomized rats model; Tryptophan hydroxylase 2; 5-hydroxytryptamine

Effects of antidepressants on osteoporosis related protein OPG, RANKL and RANK in ovariectomized rat model with depressive disorder

Objective To investigate the effects of antidepressant therapy on whole regulating function of OPG/RANKL/RANK system in ovariectomized(OVX) rat model with depressive disorder. Methods Forty sexually mature female SD rats were used as subjects. The rats were randomly divided into 8 groups as following: sham group(Sn), depression group(Dn), the OVX group(On), the OVX with depression group(ODn), OVX with depression + sertraline group (ODs), and OVX with depression + citalopram (ODx), OVX with depression + reboxetine group(ODr), OVX with depression + venlafaxine group(ODw). Bilateral oophorectomy was applied to establish the ovariectomized rats model.Chronic unpredictable mild stress(CUMS) was used to estalish depression model. Open-field test and sugar consumption experiment were applied before the experiment and the 28th day of the experiment. Immunohistochemistry and Quantitative Real-time PCR were used to observe the expression of OPG, RANKL and RANK. Results After chronic unpredictable mild stress the scores of horizontal motion, vertical motion and percentage of sugar consumption experiment in Dn group were decreased contrast to the in control Sn group (P<0.05)and the scores in ODn group were decreased in contrast to the On group (P<0.05). The protein and gene expression of RANK in Dn group and On group were increased in contrast to the Sn group (P<0.05). The protein and gene expression of RANKL in Dn group and On group were increased in contrast to the Sn group (P<0.05). The gene expression of OPG in ODs group(5.94±0.79), ODw group(5.99±1.08) and ODr group(6.59±0.69)was decreased in contrast to the ODn group(4.96±0.51)(P<0.05). The gene expression of RANK in ODs group(2.82±0.23), ODx group(3.03±0.04)was increased in contrast to the ODn group(3.44±0.18)(P<0.05). The gene expression of RANKL in ODs group(4.82±0.94), ODx group(5.00±0.82) and ODw group(5.23±0.88)was decreased in contrast to the ODn group(3.69±0.69)(P<0.05). Conclusion The expression of RANK and RANKL in femoral neck is increased both in depressive disorder model rats and ovariectomized model rats.The antidepressant can up-regulate the expression of RANK, and down-regulate the expression of OPG and RANKL in the OVX with depression group. Of the effect, sertraline is the strongest, however reboxetine mesylate is the poorest. Key words: Antidepressant; Depressive disorder rats model; Ovariectomized rats model; Osteoprotegerin; Receptor activator of NF-κB

Effects of selective serotonin and norepinephrine reuptake inhibitors on depressive- and impulsive-like behaviors and on monoamine transmission in experimental temporal lobe epilepsy.

Examine therapeutic potential of a selective serotonin reuptake inhibitor (SSRI) and a norepinephrine reuptake inhibitor (NERI) in an animal model of comorbidity between epilepsy, depression-like, and impulsive-like impairments. Epilepsy was induced in male Wistar rats by LiCl and pilocarpine. An SSRI fluoxetine (FLX), and an NERI reboxetine (RBX) were administered either alone or as a combination over 1 week. Depressive-like and impulsive-like behaviors were examined using the forced swim test. Fast scan cyclic voltammetry was used to analyze serotonergic transmission in the raphe nucleus (RN)-prefrontal cortex (PFC) pathway, and noradrenergic transmission in locus coeruleus (LC)-PFC, and LC-RN projections. Monoamine levels in PFC were measured using high-performance liquid chromatography (HPLC). Functional capacities of 5-HT1A receptors and α2A adrenoreceptors in PFC were analyzed by autoradiography. Epileptic rats showed behavioral signs of depression and hyperimpulsivity, suppressed serotonergic and noradrenergic tones, decreased levels of serotonin (5-HT), and norepinephrine (NE); 5-HT1A receptor and α2A adrenoreceptors functions remained intact. FLX failed to improve behavioral deficits, but effectively raised 5-HT level and marginally improved RN-PFC serotonergic transmission. RBX reversed impulsive-like behavior, normalized content of NE and noradrenergic tone in LC-PFC and LC-RN. FLX-RBX combination fully reversed depressive-like behavior, and normalized RN-PFC serotonergic transmission. None of the treatment modified the function of 5-HT and NE receptors. Depressive- and impulsive-like behaviors in the pilocarpine model of epilepsy stem respectively from dysfunctions of serotonergic and noradrenergic ascending pathways. At the same time, epilepsy-associated depression is SSRI resistant. The finding that an SSRI-NERI combination exerts antidepressant effect, along with RBX-induced improvement of LC-RN noradrenergic transmission point toward the involvement of LC-RN noradrenergic input in enabling therapeutic potential of FLX. Medications that improve serotonergic and noradrenergic transmission, such as serotonin-norepinephrine reuptake inhibitors may be effective in treating epilepsy-associated SSRI-resistant depression, as well as concurrent depression and attention-deficit/hyperactivity disorder (ADHD).

POP-Q indication points, Aa and Ba, involve in diagnosis and prognosis of occult stress urinary incontinence complicated with pelvic organ prolapse

To investigate the correlation between pelvic organ prolapse quantitation (POP-Q) indication points and the incidence of occult stress urinary incontinence (OSUI) and its impact on prognosis. Retrospective study medical records of 93 patients with pelvic organ prolapse (POP) staged at III-IV, of which underwent pelvic reconstruction operations with Prolift system from Jan. 2007 to Sept. 2012. None of these patients had clinical manifestations of stress urinary incontinence (SUI) before surgery, and in which 44 patients were included in study group (POP complicated with OSUI) because they were identified with OSUI, another 49 patients as control group (simple POP). Follow-up and collecting datas including POP-Q, stress test, urodynamic recordings, incidence of de novo SUI, statistic analyzing by logistic regression and receiver operating characteristic curve (ROC). (1) The study group had a much higher incidence of 30% (13/44) on de novo SUI than that of control group (4%, 2/49; P < 0.01). (2) Vaginal delivery (OR = 5.327, 95% CI: 1.120-25.347), constipation (OR = 5.789, 95% CI: 1.492-22.459), preoperative OSUI (OR = 13.695, 95% CI: 2.980-62.944), anterior vaginal wall prolapse (OR = 6.115, 95% CI: 1.231-30.379) were identified as dependent risk factors for de novo SUI by logistic regression analysis. (3) For POP patients that complicated with OSUI, we chose a cutoff value of +1.5 cm for Aa point as the threshold to predicting incidence of de novo SUI according to ROC curve, area under the curve (AUC) was 0.889 (P < 0.05), the sensitivity reached 88.9% and specificity was 73.9%. According to ROC curve of Ba point, a cutoff value of +2.5 cm was chosen as the threshold to predicting incidence of de novo SUI post-operation, it had a sensitivity of 66.7% and specificity of 82.6%, AUC was 0.766 (P < 0.05). Pre-operative OSUI is a dependent risk factor of de novo SUI for advanced POP patients. Aa and Ba points are correlated with preoperative OSUI, and it is worthy to be considered as a risk predictor on forecasting the incidence of de novo SUI post pelvic reconstruction surgery.

Antidepressant-like activity of a new piperazine derivative of xanthone in the forced swim test in mice: The involvement of serotonergic system

BackgroundThe studied compound: 3-chloro-5-{[4-(2-hydroxyethyl)piperazin-1-yl]methyl}-9H-xanthen-9-one dihydrochloride (HBK-6) is a new xanthone derivative. In this study we investigated its antidepressant-like properties and possible mechanism of action. MethodsAntidepressant-like activity was evaluated in the forced swim test (FST) in mice. The influence on locomotor activity in mice was analyzed to determine whether the observed in FST effect is specific. Rotarod test was used to determine neurotoxic properties. ResultsHBK-6 reduced immobility time in mice in FST at the doses 5 and 10mg/kg, whereas fluoxetine (FX) at 15mg/kg, reboxetine (RX) at 10mg/kg and bupropion (BPR) at 5mg/kg. Joint administration of sub-effective doses of HBK-6 and FX, but not RX or BPR, reduced immobility in mice in FST. HBK-6 at the dose 5mg/kg did not show activity in FST after pretreatment with p-chlorophenylalanine. The studied xanthone derivative at the doses 5 and 10mg/kg did not impair motor coordination in mice. ConclusionsWe demonstrated that HBK-6 has a potent antidepressant-like activity in FST, stronger than that of FX and RX, and seems to mediate its effect through serotonergic system. Moreover, at antidepressant-like doses it does not show neurotoxic properties. Given the promising results, HBK-6 may have potential in the treatment of depression, but this needs extended studies.

No evidence for enhanced extinction memory consolidation through noradrenergic reuptake inhibition—delayed memory test and reinstatement in human fMRI

One promising approach in the current ambition to maximise treatment benefit for anxiety disorders is the pharmacological enhancement of cognitive-behavioural treatment efficacy, which can be experimentally modelled by pharmacological enhancement of extinction learning/consolidation. Noradrenaline (NA) is involved in memory consolidation, and NAergic innervations are found in brain areas implicated in fear conditioning and extinction. Thus, to enhance extinction memory consolidation through boosted NAergic signalling, we administered 4mg reboxetine (RBX) immediately after extinction learning (day2, 24h after conditioning on day1) in a randomised, placebo (PLC)-controlled design. At a delayed memory test (day8), we probed cued and contextual fear and extinction memories before and after a reinstatement manipulation. After reinstatement, we find significantly enhanced amygdala and posterior hippocampus activation in the RBX group, areas implicated in fear memory expression, while the PLC group exhibited enhanced activation in areas associated with extinction memory expression (vmPFC, anterior hippocampus). No group differences were found in skin conductance responses. Thus, our data do not support our hypothesis that enhancement of NA signalling may facilitate extinction memory consolidation and provide preliminary evidence that this might rather enhance fear memories on a neural but not physiological (skin conductance responses) level.

Motor Neglect and Future Directions for Research

Motor Neglect and Future Directions for Research